US2002110840A1PendingUtilityA1

Screening method for identifying compounds that selectively induce interferon alpha

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Assignee: 3M INNOVATIVE PROPERTIES COPriority: Dec 8, 2000Filed: Dec 6, 2001Published: Aug 15, 2002
Est. expiryDec 8, 2020(expired)· nominal 20-yr term from priority
Inventors:Mark A. TomaiJohn Vasilakos
A61P 43/00A61P 7/00A61P 31/20A61P 31/10A61P 9/00A61P 31/16A61P 35/02A61P 35/00A61P 31/12A61P 31/14A61P 37/04A61P 25/28A61P 19/08G01N 33/5047G01N 2333/54A61P 19/04G01N 2333/56G01N 2333/525A61P 17/02G01N 33/6866A61P 21/00G01N 33/6863A61P 17/04G01N 2500/20A61P 17/00A61P 1/16
46
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Claims

Abstract

Methods for screening for compounds that selectively induce IFN-&60 production and methods for ameliorating conditions in a patient using a small molecule that selectively induces the production of IFN-α are disclosed.

Claims

exact text as granted — not AI-modified
We claim:  
     
         1 . A method for identifying a compound that selectively induces production of IFN-α from pDC2 cells, the method comprising: 
 obtaining a population of cells that includes both inflammatory cytokine producing cells and pDC2 cells;  
 contacting the population of cells with a test compound;  
 determining the amount of IFN-α present in the population of cells contacted with the test compound;  
 determining the amount of inflammatory cytokine(s) present in the population of cells contacted with the test compound; and  
 identifying the test compound as a selective inducer of IFN-α if IFN-α is present in the population of cells after contact with the test compound in an amount at least three times greater than the amount of inflammatory cytokine(s) present in the population of cells.  
 
     
     
         2 . The method of  claim 1  wherein the amount of IFN-α and inflammatory cytokine(s) is determined from culture supernatants using ELISA or bioassay.  
     
     
         3 . The method of  claim 1  wherein the amount of IFN-α and inflammatory cytokine(s) is determined from cells in the population using a method selected from the group consisting of dot blotting, Western blotting, Northern blotting, RPA and RT-PCR.  
     
     
         4 . The method of  claim 1  wherein the inflammatory cytokine is TNF-α or IL-12.  
     
     
         5 . The method of  claim 1 , wherein the population of cells is in whole blood.  
     
     
         6 . The method of  claim 1 , wherein the population of cells comprises peripheral blood mononuclear cells.  
     
     
         7 . The method of  claim 1 , wherein the population of cells comprises a fraction of peripheral blood mononuclear cells containing at least 5% pDC cells.  
     
     
         8 . The method of  claim 1  wherein the population of cells is contacted with the test compound at concentrations ranging from about 0.005 to 5 μM.  
     
     
         9 . The method of  claim 1  wherein the population of cells is cultured with the test compound for a period ranging from about 12 to 36 hours.  
     
     
         10 . The method of  claim 1  wherein the population of cells comprises a CD14+ cell type.  
     
     
         11 . The method of  claim 10  wherein the inflammatory cytokine is TNF-α and the amount of TNF-α produced by the CD14+ cells is undetectable when the test compound contacts the population of cells at a concentration of 1 μM.  
     
     
         12 . A method for identifying a compound that selectively induces production of IFN-α from pDC2 cells, the method comprising: 
 obtaining a population of cells that includes both inflammatory cytokine producing cells and pDC2 cells;  
 contacting the population of cells with a test compound;  
 identifying pDC2 cells present in the population and determining that IFN-α is produced by the pDC2 cells by flow cytometry;  
 determining the production of inflammatory cytokine(s) in the population of cells by flow cytometry; and  
 identifying the test compound as a selective inducer of IFN-α if all cells present in the population other than pDC2 cells produce insignificant levels of inflammatory cytokine(s).  
 
     
     
         13 . The method of  claim 12  wherein the pDC2 cells are identified by the presence of HLA-DR and CD123 cell surface markers on a surface of the pDC2 cells.  
     
     
         14 . The method of  claim 12  wherein the inflammatory cytokine is TNF-α, IL-12 and/or IL-1.  
     
     
         15 . The method of  claim 12 , wherein the population of cells is in whole blood.  
     
     
         16 . The method of  claim 12 , wherein the population of cells comprises peripheral blood mononuclear cells.  
     
     
         17 . The method of  claim 12 , wherein the population of cells comprises a fraction of peripheral blood mononuclear cells.  
     
     
         18 . The method of  claim 12  wherein the population of cells is contacted with the test compound at concentrations ranging from about 0.005 to 5 μM.  
     
     
         19 . The method of  claim 12  wherein the population of cells is cultured with the compound for a period ranging from about 2 to 24 hours.  
     
     
         20 . A method of affecting a condition of a patient responsive to IFN-α, the method comprising a step of: 
 obtaining a population of cells that includes both inflammatory cytokine producing cells and pDC2 cells;  
 contacting the population of cells with a test compound;  
 determining the amount of IFN-α present in the population of cells contacted with the test compound;  
 determining the amount of inflammatory cytokine(s) present in the population of cells contacted with the test compound;  
 identifying the test compound as a selective inducer of IFN-α if IFN-α is present in the population of cells after contact with the test compound in an amount at least three times greater than the amount of the inflammatory cytokine(s) present in the population of cells; and  
 administering the identified selective compound to the patient to affect the condition.  
 
     
     
         21 . The method of  claim 20  wherein the amount of IFN-α and inflammatory cytokines is determined from culture supernatants using ELISA or bioassay.  
     
     
         22 . The method of  claim 20  wherein the amount of IFN-α and inflammatory cytokines is determined from cells in the population using a method selected from the group consisting of dot blotting, Western blotting, Northern blotting, RPA and RT-PCR.  
     
     
         23 . The method of  claim 20  wherein the inflammatory cytokines include TNF-α and/or IL-1.  
     
     
         24 . The method of  claim 20 , wherein the population of cells is in whole blood.  
     
     
         25 . The method of  claim 20 , wherein the population of cells comprises peripheral blood mononuclear cells.  
     
     
         26 . The method of  claim 20 , wherein the population of cells comprises a fraction of peripheral blood mononuclear cells.  
     
     
         27 . The method of  claim 20  wherein the population of cells is contacted with the test compound at concentrations ranging from about 0.005 to 5 μM.  
     
     
         28 . The method of  claim 20  wherein the population of cells are cultured with the test compound for a period ranging from about 12 to 36 hours.  
     
     
         29 . The method of  claim 20  wherein the population of cells comprises a CD14 + or DC1 cell type.  
     
     
         30 . The method of  claim 29  wherein the inflammatory cytokine is TNF-α and the amount of TNF-α produced by the CD14 + or DC1 cells is undetectable when the test compound is in contact with the cells at a concentration of about 1 μM.  
     
     
         31 . The method of  claim 20  wherein the condition is selected from the group consisting of melanoma, myeloid leukemia, non-Hodgkin's lymphoma, renal cell carcinoma, Kaposi's sarcoma, multiple sclerosis, hypereosinophilic syndrome, myoproliferative disorders, idiopathic myelofibrosis, hepatitis B, and chronic hepatitis C, variola, influenza, parainfluenza, adenovirus, coronavirus, and rhinovirus.  
     
     
         32 . The method of  claim 31  wherein the condition is selected from the group constiting of cutaneous necrotising vasiculitis, mixed cryoglobulinemia, porphyria cutanea tarda, lichen planus, Adamantiadis-Behcet syndrome, erythema multiforme and nodosum, malacoplakia, urticaria and pruritus.  
     
     
         33 . The method of  claim 20  wherein the selective compound is delivered to a desired cell type by a targeting moiety.  
     
     
         34 . A method for selectively inducing IFN-α production from a population of cells that includes pDC2 cells, the method comprising contacting the population of cells with an immune response modifier compound that selectively induces IFN-α production by pDC2 cells.  
     
     
         35 . The method of  claim 34  wherein the compound is Compound I.  
     
     
         36 . The method of  claim 34  wherein the compound is Compound II.  
     
     
         37 . The method of  claim 34  wherein the compound is Compound III.  
     
     
         38 . The method of  claim 34  wherein the compound is Compound IV.  
     
     
         39 . The method of  claim 34  wherein the compound is Compound V.  
     
     
         40 . The method of  claim 34  wherein the compound is Compound VI.

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